what passage is too high for transient transfection? - (Dec/02/2008 )

I'd like to transfect rhabdomyosarcoma cell line with a plasmid for detecting luciferase activity. They're currently already at passage 25, which I know is not optimal for transfection efficiency, but when is it too high to even bother trying? I will probably be using SuperFect from Qiagen (or if someone has a better recommendation).

Thanks so much!

---

It has been taught me early passages are the choices for good transfection. However, your cells are enough good, new stock (passaged only 2-3 times i.e. stock passage was 23), I think only higher passage like 25 will not bother your trasfection efficiency.

Selection of transfection agent and amount, cell density and cell type, cell viability are more important factors to be cared. If you have optimized protocol with your cells and transfection agent, then you will surely get a high efficiency of transfection.

I do not have any idea on myosarcoma cells. I have been mostly using Lipofectamine and GeneJammer for epithelial and fibrosarcoma cells respectively.

Good luck.

---